Chemotaxonomy of four varieties of Mentha longifolia L. using essential oil composition markers

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J. Bio. & Env. Sci. 2014 Introduction Mentha longifolia L. is a cosmopolitan genus of over specious, belonging to Mentha tribe, Lamiaceae family(Mozaffarian, 1999). This genus comprises about 25specious in Iran which 7 of them endemic (Mozaffarian, 1999). Mentha longifolia L. Huds. Is one of the specious of this genus that grows as wild in various regions of Iran (Lange and Croteau, 1999). This species is not only used in food processing technology as flavors or specious but also in hygiene and pharmaceutical products because of it`s antimicrobial properties. It is used in Iranian traditional medicines as a stomach pain- relieving agent, antispasmodic, digestive and carminative (Zargari, 1990). This specious show a large phenotypic plasticity and most varieties of it are capable of hybridization with each other. It has six varieties in Iran (Mozaffarian, 1999). The composition of volatile compounds is known for this species. (Abbaszadeh et al., 2013, Hajlaoue et al., 2010, Saeidi et al., 2012, Sharpov et al., 2011), and some data are found in chemotaxo-nomy studies of this species. Chemotaxonomy of mints of genus Mentha by applying Raman spectroscopy (Rosch et al.,2002). Essential oil of Five Mentha species using as-liquid partition chromatography was studied (Handa et al.,2006). Isopiperitenone and terpenoids were detected the main constituents between other components and correlation of the chemical data illustrates their value in plant classification. Also the chemotypes of M. spicata was studied in Greece and their chemotypes were characterized (Kokkini et al.,1989). M. longifolia exhibits great morphological and chemical variability. Therefore, the present study aims to investigate the volatile compounds of four varieties of this species, and evaluate the chemotaxonomic relationships of these four varieties of this specious. Material and methods Plant material Five specious from four varieties of M. longifolia were collected during the flowering period from Tehran province and Northera parts of country (table 1). Table 1. Varieties of M. longifolia and their localities. Variety Locality M. longifolia var. callianta Firoozkohroad to Damavand, 2166m, Mazooji M. longifolia var. chlorodictya Gilan: Asalem to Khalkhal, 1963m, Mazooji M. longifolia var. longifolia Tehran: Firoozkohroad,Chapdare,2134m,Mazooji M. longifolia var. aciatica Mazandaran: Haraz road to lar,2270m,Mazooji M. longifolia var. chlorodictya Gilan: Asalem to Khalkhal, 1963m, Mazooji The taxonomic description of these taxa, follows flora Iranica. Essential oil extraction For isolation of the volatile oils, the aerial parts of the plants were dried at room temperature and hydrodistilled for 5h anhydrous sodium thiosulfate and kept at 4°c in sealed required. Analytical gas chromatography was capillary column DB-5 (30 m. 0.25 mmid, 0.25 µm film thickness); Carrier gas, He; Split ratio, 1: 25°, and using a flame ionization detector. The column temperature was programmed at 50°c for 1 min, and then heated to 265°c at a rate of 2.5°c/min, then kept constant at 265°c for 20 min. GC-MS was performed on a thermoquest 2000 with quadruple detector, on capillary column DB-5(GC), carried gas, He; Flow rate, 1.5 ml/min. The column was held at 50°c for 1 min, and programmed up to 265°c for 20 min. Quantitative data were obtained from the electronic integration of the FID peak areas. Cluster analysis Hierachial clustering analysis of main components between species was performed using SPSS software 173 | Barzin et al.
J. Bio. & Env. Sci. 2014 with Average Linkage method and the resulting dendrogram was illustrated. Result and discussion The classification of aromatic lants considered to belong to the Mentha- an important source of basic raw materials for the food, drug and cosetic industries- has always proved a difficult taske to the botanists and geneticist. Based on our results,Fifty one compounds were detected in the oil of four varieties representing the 96-74 to 98-52% of the total oil. The major constituents were Carvon, 1,8- cineol, cis-piperitone oxide, Pulegone, Menthone and iso-Menthone. The most chemical components showed two main clades (Fig 1). One main clade consists of M. longifolia var. chlorodictya 2 as a separate clade with highest distance from the others varieties. The second main clade consist of four other varieties. As shown in Fig 1, in this clade, M. longifolia var. aciatica formed one subclade in 21 taxonomic level and three other specimens are placed in another subclade. There are very similarities between two varieties consist of M. longifolia var. chlorodictya 2 . The PCA confirmed the cluster analysis results (Fig 2). Carvone –Pulegone ratios, were shown to distinguish readily two similar varieties consist of M. longifolia var. longifolia and M. longifolia var. chlorodyctia². These two varities placed in one subclade near each other (Fig 1). The occurence of Menthon-Pulegone in M. longifolia var.asiatica M.longifolia var. callicantha. But in M. longifolia var. chlorodictia ¹, major constituent are Iso-Menthone and Pulegone and can distict this variety from others. Fig. 1. Cluster analysis of four varieties resulted from analysis of chemical components data (Abbreviation: 1- M. longifolia var. longifolia , 2- M. longifolia var. chlorodictya 1 , 3- M. longifolia var.asiatica, 4- M.longifolia var. chlorodictya 2 , 5- M. longifolia var. callianta). 174 | Barzin et al.
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Chemotaxonomy of four varieties of Mentha longifolia L. using essential oil composition markers

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